振动对体外血管内皮细胞中炎症因子及IncRNA MEG3表达的影响
Effects of vibration on expressions of vascular endothelial inflammatory factors and IncRNA MEG3 in vitro
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摘要:
背景 长时间暴露于振动可导致血管内皮损伤，炎症反应在血管内皮损伤中起着重要作用。研究表明长链非编码RNA（lncRNA）母系表达基因3（
MEG3 ）参与内皮细胞的炎症损伤。目的 探究振动对体外血管内皮细胞分泌炎症因子及IncRNA
MEG3 表达的影响。方法 将人脐静脉内皮细胞（HUVEC）分为振动组和对照组：振动组分别施加63 Hz、6.76 m·s−2，200 Hz、5.08 m·s−2，250 Hz、4.56 m·s−2频段的振动，每日接振1~4 h，分别暴露1、2 d；对照组除不接触振动外，其他均与实验组相同。采用CCK-8检测不同振动频率和时间对HUVEC活力的影响。采用酶联免疫吸附试验检测细胞和上清液中肿瘤坏死因子-α（TNF-α）、白介素（IL）-8、IL-4、IL-10的浓度。通过实时荧光定量PCR检测IncRNA
MEG3 的表达水平。结果 与对照组相比，63 Hz振动1 d组，1.5、3 h后细胞活力上升，2、2.5 h后下降；2 d组，振动1.5 h后细胞活力上升，2、2.5 h后下降。200 Hz振动1 d组，振动2、4 h后细胞活力上升，2.5、3 h后下降；2 d组，振动1.5 h后细胞活力上升，2.5 h后下降。250 Hz振动1 d组，1.5、2.5 h后细胞活力上升，3 h后下降；2 d组，振动1.5 h后细胞活力上升，3 h后下降。在63、200 Hz振动2.5 h，250 Hz振动3 h条件下，与对照组相比：1 d组和2 d组TNF-α、IL-8、IL-4、IL-10在细胞和上清液中浓度均升高；1 d组lncRNA
MEG3 的表达水平下降，2 d组仅63 Hz振动后lncRNAMEG3 的表达水平下降。以上结果均具有统计学意义（P <0.05）。结论 振动可以诱导体外血管内皮细胞中炎症因子TNF-α、IL-8、IL-4、IL-10水平升高以及lncRNA
MEG3 表达水平下降。Abstract:Background Prolonged exposure to vibration can cause vascular endothelial injury, and inflammatory response plays an important role in vascular endothelial injury. Studies have shown that long non-coding RNA (lncRNA) maternally expressed gene 3 (
MEG3 ) is involved in regulating the expression of inflammatory injury of endothelial cells.Objective To investigate the effects of vibration on the secretion of inflammatory factors and the expression of IncRNA
MEG3 by vascular endothelial cellsin vitro .Methods Human umbilical vein endothelial cells (HUVEC) were divided into two categories: vibration and control. The vibration exposure included 63 Hz (6.76 m·s−2), 200 Hz (5.08 m·s−2), and 250 Hz (4.56 m·s−2) frequency bands, and 1 and 2 d exposure time with 1 to 4 h of daily vibration. The control treatment was the same as the vibration category except that they were not exposed to vibration. CCK-8 was used to detect the effects of different vibration frequencies and time on the viability of HUVEC. The expression levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-4 (IL-4), and interleukin-10 (IL-10) in the cells and supernatants were detected by enzyme-linked immunosorbent assay. The expression levels of IncRNA
MEG3 were detected by real-time fluorescence quantitative PCR.Results Compared with the cells with the control treatment, the cell viability of the 1-day exposure group increased after 1.5 h and 3 h of vibration at 63 Hz, while decreased after 2 h and 2.5 h; the cell viability of the 2-day exposure group increased at the frequency of 63 Hz for 1.5 h, but decreased at 2 h and 2.5 h. At the frequency of 200 Hz, the cell viability of the 1-day exposure group increased at 2 h and 4 h, but decreased at 2.5 h and 3 h; the cell viability of the 2-day exposure group increased at 1.5 h and decreased at 2.5 h. For the vibration exposure at frequency of 250 Hz, the cell viability of the 1-day exposure group increased at 1.5 h and 2.5 h, but decreased at 3 h; of the 2-day exposure group, the cell viability increased at 1.5 h and decreased at 3 h. For the exposure settings of 63 and 200 Hz vibration for 2.5 h and 250 Hz vibration for 3 h, and with the control treatment as reference, the expression levels of TNF-α, IL-8, IL-4, and IL-10 in cells and supernatants were increased in the 1 d and 2 d exposures; the expression level of lncRNA
MEG3 decreased in the 1 d exposure group; however, for the 2 d exposure, the expression level of lncRNAMEG3 decreased only in the 63 Hz vibration exposure. All of these results were statistically significant (P <0.05).Conclusion Vibration could induce an increase in the levels of inflammatory factors TNF-α, IL-8, IL-4, and IL-10 and a decrease in the expression level of lncRNA
MEG3 in vascular endothelial cellsin vitro .
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